D'Agostino, Ylenia (0007) Generation and characterization of BDNF knock-out in zebrafish by CRISPR/Cas9 system. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Lingua: English
Title: Generation and characterization of BDNF knock-out in zebrafish by CRISPR/Cas9 system
Creators:
CreatorsEmail
D'Agostino, Yleniayleniadag87@gmail.com
Date: 17 April 0007
Number of Pages: 196
Institution: Università degli Studi di Napoli Federico II
Department: Medicina Veterinaria e Produzioni Animali
Dottorato: Biologia
Ciclo di dottorato: 29
Coordinatore del Corso di dottorato:
nomeemail
Cringoli, Giuseppecringoli@unina.it
Tutor:
nomeemail
D'Aniello, SalvatoreUNSPECIFIED
Date: 17 April 0007
Number of Pages: 196
Uncontrolled Keywords: BDNF, Neurotrophin, CRISPR/Cas9, genome-editing, knock-out, zebrafish
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/11 - Biologia molecolare
Area 05 - Scienze biologiche > BIO/12 - Biochimica clinica e biologia molecolare clinica
Area 05 - Scienze biologiche > BIO/13 - Biologia applicata
Area 05 - Scienze biologiche > BIO/18 - Genetica
Additional Information: Attività di ricerca svolta presso i laboratori della Stazione Zoologica Anton Dohrn di Napoli
Date Deposited: 03 May 2017 08:18
Last Modified: 07 Mar 2018 09:38
URI: http://www.fedoa.unina.it/id/eprint/11576

Abstract

BDNF is one of the main neurotrophin (NT) expressed in the brain as well in some peripheral sensory neurons. The important role played by BDNF is demonstrated by its well-established effects on axon growth, synaptic plasticity and, not less important, in processes of learning and memory. Moreover, BDNF is becoming a valid candidate for treatment of neurodegenerative disorders such as epileptic seizures, anxiety and depression or Alzheimer's and Parkinson's diseases. However, unless the huge number of studies performed about its role, some points remain controversial and the whole scenario is still far from be fully delineated. Most of the information we currently have about BDNF function derive from studies in mice but, in this case, the major obstacle is represented by the early postnatal lethality of BDNF-/- and, as consequence, all the results are limited to BDNF+/- animals. The main objective of this PhD thesis was the generation of a BDNF knock-out line in zebrafish through the CRISPR/Cas9 technology and the characterization of the mutant phenotype from both molecular and behavioural points of view. My results show that zebrafish BDNF-/-, unlike from what happens in mouse, survive after birth with undetectable levels of BDNF protein and does not present any mutation in undesirable off-target gene. Behavioural test through larvae monitoring reveals a significantly lower activity, suggestive of reduced exploratory behaviour, while several anomalies such as aberrant pharyngeal arches formation or increasing of apoptotic cells underline the complex role played by BDNF and its implication in a variety of biological processes. Differential transcriptomic analysis between mutant and wild type has been also performed in order to identify downstream target gene regulated by BDNF. The ongoing analysis of the RNA-Seq data will provide new insights on the role played by BDNF during the development of the nervous system. Finally, the zebrafish knock-out line also provides a promising tool in terms of diagnosis and treatment of neurodegenerative diseases in which this important NT is implicated.

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